Gene Therapy of Human Hemophilia
Faculty Information
Name:
Prof. Arun Srivastava
Email
aruns@ufl.edu
Phone
(352) 273-8259
Faculty Department/Division
Molecular Genetics & Microbiology
This project is primarily:
Basic
Research Project Description:
Several first generation of AAV serotype vectors (AAV2, AAV5, AAV6, AAV8, AAVrh10, and AAVrh74) vectors, composed of naturally occurring capsids, that have been used for gene therapy of hemophilia B, but none of these serotype vectors are truly human liver-tropic. We first identified AAV3 vectors for their remarkable tropism for primary human hepatocytes. We have also generated capsid-modified next generation (NextGen), and genome-modified generation X (GenX, and GenY), and the combination of the two, optimized (Opt AAV3) vectors. We have also documented the safety and remarkable specificity and efficacy of one of these novel AAV3 vectors in “humanized” mice and in non-human primate models. NextGen AAV3 vectors have been shown to perform significantly better than the first generation AAV2, AAV5, AAV6 and AAV8, and AAVrh74 vectors in expressing the therapeutic levels of human clotting factor IX (hFIX) gene in “humanized” mice and in non-human primates at significantly lower vector doses. Similarly, gene therapy of hemophilia A has also been performed with several AAV serotype vectors (AAV5, AAV-LK03, and AAV6) expressing the human clotting factor VIII (hVIII) for gene therapy of hemophilia, but none has proven to be optimal since all these vectors target hepatocytes for transgene expression. FVIII is normally made in liver sinusoidal endothelial cells (LSECs), which are able to efficiently secrete FVIII. None of the aforementioned AAV capsids has been shown to efficiently target human LSECs. We have documented that AAV3 vectors also transduce human LSECs most efficiently than AAV5 and AAV6 vectors. Taken together, our data suggest that AAV3 vectors may also prove to be efficacious as well as cost-effective in gene therapy for hemophilia A. Medical students will be involved in evaluating the efficacy of further optimized AAV3 vectors in primary human hepatocytes and in LSECs, respectively, for the potentially safe and efficacious gene therapy of hemophilia B and hemophilia A. Funding for the project is provided by the National Institutes of Health and the George H. Kitzman Endowment.
D. Markusic, R.W. Herzog, G. Aslanidi, B. Hoffman, B. Li, M. Li, G.R. Jayandharan, C. Ling, I. Zolotukhin, W. Ma, S. Zolotukhin, A. Srivastava, and L. Zhong. High-efficiency transduction and correction of murine hemophilia B using AAV2 vectors devoid of multiple surface-exposed tyrosines. Mol. Therapy, 18: 2048-2056, 2010.
D. Markusic, T.C. Nichols E.P. Merricks, B. Palaschak, I. Zolotukhin; D. Marsic; S. Zolotukhin, A. Srivastava, and R.W. Herzog. Evaluation of engineered AAV capsids for hepatic factor IX gene transfer in murine and canine models. Journal of Translation Medicine, 15: 94, 2017.
H.C. Brown, C.B. Doering, R.W. Herzog, C. Ling, D.M. Markusic, H.T. Spencer, A. Srivastava, and A. Srivastava. Development of a clinical candidate AAV3 vector for gene therapy of hemophilia B. Human Gene Therapy, 31: 1114-1123, 2020.
S.R.P. Kumar, J. Xie, S. Hu, J. Ko, Q. Huang, H.C. Brown, A. Srivastava, D.M. Markusic, C.B. Doering, H.T. Spencer, A. Srivastava, G. Gao, and R.W. Herzog. Coagulation factor IX gene transfer to non-human primates using engineered AAV3 capsid and hepatic optimized expression cassette. Molecular Therapy Methods & Clinical Development, 23: 98-107, 2021.
Shoti, K. Qing, and A. Srivastava. Development of an AAV DNA-based synthetic vector for the potential gene therapy of hemophilia in children. Frontiers in Microbiology, 13: 1033615, 2022.
Does this project have an international component or travel?
No